Hi,

You should purify both DNA fragments via gel extraction if the insert originates from another plasmid. Afterwards, you can try different concentration ratios of both DNA molecules in ligation mixes. Starting with a 3:1 ratio of insert:backbone would be a good idea. The ligation should be carried out over night at a temperature specified by your ligation supplier (often 4-12 C). Afterwards, you transform the ligation mix into competent E. coli cells via heat shock or electroporation. Selection for the appropriate resistance is not enough to find the desired clone. Some original pet32a molecules will be transformed along with your ligation mix. Therefore, you should screen the colonies via colony PCR. Finally, the insert sequence should be confirmed by sequencing.

Do you have some more specific questions concerning the procedure?

Good luck!

Thanks  a lot

Boas is right . Another way is TA cloning. Goodluck